Cell growth-promoting peptide and use thereof

Active Publication Date: 2014-11-13
TOAGOSEI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patent describes a cell growth promoter that can be used to promote cell proliferation without the need for expensive growth factors like bFGF. The promoter contains a peptide that can be easily produced using chemical synthesis, which reduces costs and allows for the efficient repair or regeneration of affected areas in patients. The peptide is designed to have a specific amino acid sequence and a short peptide chain, which further reduces costs and allows for ease of handling and chemical synthesis. Overall, this patent provides a cost-effective and efficient solution for promoting cell growth for research and therapeutic purposes.

Problems solved by technology

In the field of regenerative medicine, it has been a challenge to establish a method to proliferate cells of interest at a higher rate.
However, as the currently available bFGF is very expensive, it is financially difficult to use the growth factor in a relatively large quantity for cell proliferation.
Moreover, using bFGF for the purpose of cell proliferation may become a significant cause to increase the cost of cell manufacturing and tissue regeneration involving the said proliferation.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Peptide Synthesis

[0158]A total of 21 peptides (samples 1 to 21) were prepared using the subsequently described peptide synthesizer. Data on these synthesized peptides, e.g., the amino acid sequence and so forth, is represented by Tables 1 and 2.

TABLE 1total number sampleamino acidof aminono.sequenceacid residues1MLPSLALLLLAAWTVR31AGKKRTLRKNDRKKR(SEQ ID NO: 21)2PSLALLLLAAWTVRAG29KKRTLRKNDRKKR(SEQ ID NO: 22)3SLALLLLAAWTVRAGK28KRTLRKNDRKKR(SEQ ID NO: 23)4LALLLLAAWTVRAGKK27RTLRKNDRKKR(SEQ ID NO: 24)5ALLLLAAWTVRAGKKR26TLRKNDRKKR(SEQ ID NO: 25)6LLLLAAWTVRAGKKRT25LRKNDRKKR(SEQ ID NO: 26)7LLLAAWTVRAGKKRTL24RKNDRKKR(SEQ ID NO: 27)8LLAAWTVRAGKKRTLR23KNDRKKR(SEQ ID NO: 28)9LAAWTVRAGKKRTLRK22NDRKKR(SEQ ID NO: 29)10AAWTVRAGKKRTLRKN21DRKKR(SEQ ID NO: 30)11AWTVRAGKKRTLRKND20RKKR(SEQ ID NO: 31)12WTVRAGKKRTLRKNDR19KKR(SEQ ID NO: 32)

TABLE 2totalnumber ofsampleamino acidamino acidno.sequenceresidues13MLPSLALLLLAAWTV29GKKRTLRKNDRKKR(SEQ ID NO: 33)14MLPSLALLLLAAWTG28KKRTLRKNDRKKR(SEQ ID NO: 34)15MLPSLAL...

example 2

Evaluation of the Cell Growth-Promoting Activity of the Synthetic Peptides

[0183]Experimental sections were set up that used the cell growth-promoting peptides obtained in Example 1 (samples 1 to 21) as a cell growth promoter and that used a commercially available bFGF as a cell growth promoter as a comparative example. For the control, a peptide-free section (the bFGF was also not added) was set up.

[0184]The details of the evaluation testing are provided below. Each synthesized sample peptide was dissolved in PBS (phosphate-buffered physiological saline) to prepare a stock solution with a peptide concentration of 1 mM.

[0185]Rat bone marrow-derived stem cells (mesenchymal stem cells) were used as the test cells. Specifically, the test stem cells were cultured by passage on Dulbecco's MEM medium (DMEM medium: Gibco product) containing 10% fetal bovine serum (FBS: Gibco product), 2 mM of L-glutamine, 50 unit / mL of penicillin, and 50 μg / mL of streptomycin. From the second passage to the...

example 3

Granule Preparation

[0193]50 mg of the sample 1 peptide was mixed with 50 mg crystalline cellulose and 400 mg lactose; 1 mL of a mixed solution of ethanol and water was added; and mixing / kneading was carried out. The resulting mixture was granulated by a standard method to obtain granules (i.e., a granular cell growth promoter) in which the base component was the cell growth-promoting peptide.

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Abstract

The pharmaceutical composition provided by the present invention comprises at least one pharmaceutically acceptable carrier, and an active ingredient including an artificially synthesized peptide comprises: (A) an amino acid sequence constituting a cell-penetrating peptide and (B) an amino acid sequence constituting the signal peptide in amyloid precursor protein (APP) or an N-terminal partial amino acid sequence or C-terminal partial amino acid sequence from the amino acid sequence constituting that signal peptide.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application is a Continuation-in-Part of application Ser. No. 13 / 701,747 filed on Dec. 3, 2012, which is a Section 371 National Phase of PCT International application No. PCT / JP2011 / 062809, the entire contents of both of which are hereby incorporated herein by reference. This application also claims priority right based on Japanese Patent Application No. 2010-128648 filed on Jun. 4, 2010, which is hereby incorporated by reference.TECHNICAL FIELD[0002]The present invention relates to a peptide capable of promoting proliferation of stem and other cells; and use thereof. Especially, it relates to a cell growth promoter (a composition) containing the peptide and a method to produce cells of interest at an increased growth rate using the peptide.BACKGROUND ART[0003]In the field of regenerative medicine, it has been a challenge to establish a method to proliferate cells of interest at a higher rate. In the fields of cell engineering and fe...

Claims

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Application Information

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IPC IPC(8): C12N5/074
CPCC12N5/0607C12N2501/998C12N5/0663C07K14/4711C07K2319/74
InventorYOSHIDA, TETSUHIKOKOBAYASHI, NAHOKO
OwnerTOAGOSEI CO LTD